CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



695 



(1) Dissolve 3.0 g. of agar in 100.0 cc. 

 Ringer's solution. 



(2) Add a drop of dog blood to 2 to 3.0 

 cc. of dog serum diluted with 2 to 5 

 parts Ringer's solution. (Sera or 

 ascitic fluids of other animals may 

 be used.) 



(3) To 9.0 cc. of (2) add 1.0 cc. of (1). 



(4) Heat at 56 to 58°C. in a water bath 

 for about 30 minutes. 



(5) Cover with a layer of liquid sterile 

 paraffin. 



(b) Stitt cultivated Leptospira icteroides 

 (cause of yellow fever) on a medium 

 prepared as follows: 



(1) Mix one part rabbit s erum and 3 

 parts of Ringer's solution solidified 

 partially by the addition of 0.3% 

 agar. 



(2) Tube in tall tubes. 



(3) Introduce 1.0 cc. of yellow fever 

 patients blood into the lower part 

 of each tube. 



(4) Pour a thin layer of petroleum over 

 the top of the medium. 



References: Abbott (1921 p. 635), Kaneko 

 (1921-22 p. 354), Stitt (1923 p. 54). 



2154. Tanner's Serous Fluid Agar 



Constituents : 



1. Distilled water 100.0 cc. 



2. Agar 1-5 g. 



3. Serum 100.0 cc. 



Preparation: 



(1) Dissolve 1.5 g. agar in 100.0 cc. dis- 

 tilled water. 



(2) Filter and tube in 5.0 cc. quantities. 



(3) Add an equal volume of sterile serum 

 to each tube of sterile (2) melted and 

 cooled to 40°C. 



(4) Roll the tube to mix. 



(5) Cool in a slanting position. 

 Sterilization: Method of sterilization of 



(2) not given. 

 Use: General culture medium. Teague 



and Deibert used a similar medium to 



study growth requirements of Unna- 



Ducrey bacillus. 

 Variants : 



(a) Harvey specified that the agar be 

 sterilized at 120°C., and be mixed 

 with an equal volume of sterile serum 

 or ascitic fluid. 



(b) Teague and Deibert reported that the 

 Unna-Ducrey bacillus would not 

 grow on a medium prepared by mix- 

 ing various amounts of sheep or 

 rabbit serum, heated at 55°C. for 15- 

 minutes with a 2.0% agar solution 

 containing 0.5% NaCl. 

 References: Tanner (1919 p. 70), Harvey 



(1921-22 p. 79), Teague and Deibert 



(1922 p. 70). 



2155. Kanthack and Stephens' Serous 

 Exudate Agar 



Constituents: 



1. Serous exudate (ser- 

 um, ascitic fluid, etc.). 100.0 cc. 



2. KOH 10.0% 2.0 cc. 



3. Agar (1.5 to 2.0%) . . . 15.0 to 20.0 g. 



4. Glycerol (4.0 to 



5.0%) 4.0 to 5.0 cc. 



Preparation : 



(1) To 100.0 cc. of a serous exudate add 

 2.0 cc. of 10% KOH solution. (It is 

 always best to first heat a small 

 portion of the serous exudate in a 

 test tube. If it solidifies or if a large 

 amount of albumin appears, add at 

 least a double volume of distilled 

 water before adding KOH.) 



(2) Soak agar in acidified water and wash 

 free from acid. 



(3) Add 1.5 to 2.0% agar to (1). 



(4) Boil until the agar is completely 

 dissolved. (The Mixture appears 

 clear when agar is dissolved.) 



(5) Filter thru a coarse filter paper using 

 a hot water funnel. 



(6) Add 4.0 to 5.0% glycerol. 



(7) Distribute into test tubes. 

 Sterilization : Not specified. 



Use : Diagnosis of diphtheria. The authors 

 reported that staphylococci and B. coli 

 communis, etc., development was in- 

 hibited. The sooner the exudate was 

 used, after it left the human body the 

 clearer the medium. 



Variants : 



(a) The authors reported that the addi- 

 tion of 2.0 to 4.0% glucose did not 

 improve the medium. 



(b) Harvey prepared a similar medium as 

 follows: 



(1) Collect ascitic fluid with sterile 

 precautions. 



